Cellular context determines primary characteristics of human TRPC5 as a cold-activated channel

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F67985823%3A_____%2F22%3A00561637" target="_blank" >RIV/67985823:_____/22:00561637 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/00216208:11310/22:10452947

Výsledek na webu

<a href="https://doi.org/10.1002/jcp.30821" target="_blank" >https://doi.org/10.1002/jcp.30821</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1002/jcp.30821" target="_blank" >10.1002/jcp.30821</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Cellular context determines primary characteristics of human TRPC5 as a cold-activated channel

Popis výsledku v původním jazyce

The human transient receptor potential canonical 5 (TRPC5) is a calcium-permeable, nonselective cation channel expressed in the central and peripheral nervous system and also in other tissues such as the kidney, synovium, and odontoblasts. TRPC5 has been recently confirmed to play a key role in spontaneous, inflammatory mechanical, and cold pain. Although TRPC5 activation is known to be cold sensitive, it is unclear whether this property is intrinsic to the channel protein and whether or to what extent it may be determined by the cellular environment. In this study, we explored the cold sensitivity of human TRPC5 at the single-channel level using transiently transfected HEK293T cells. Upon decreasing the temperature, the channel demonstrated prolonged mean open dwell times and a robust increase in the open probability (P-o), whereas the amplitude of unitary currents decreased similar to 1.5-fold per 10 degrees C of temperature difference. In the absence of any agonists, the temperature dependence of P-o was sigmoidal, with a steep slope within the temperature range of 16 degrees C-11 degrees C, and exhibited saturation below 8-5 degrees C. Thermodynamic analysis revealed significant changes in enthalpy and entropy, suggesting that substantial conformational changes accompany cold-induced gating. The mutant channel T970A, in which the regulation downstream of G-protein coupled receptor signaling was abrogated, exhibited higher basal activity at room temperature and a less steep temperature response profile, with an apparent threshold below 22 degrees C. An even more pronounced decrease in the activation threshold was observed in a mutant that disrupted the electrostatic interaction of TRPC5 with the endoplasmic reticulum calcium sensor stromal interaction molecule 1. Thus, TRPC5 exhibits features of an intrinsically cold-gated channel, its sensitivity to cold tightly depends on the phosphorylation status of the protein and intracellular calcium homeostasis.

Název v anglickém jazyce

Cellular context determines primary characteristics of human TRPC5 as a cold-activated channel

Popis výsledku anglicky

The human transient receptor potential canonical 5 (TRPC5) is a calcium-permeable, nonselective cation channel expressed in the central and peripheral nervous system and also in other tissues such as the kidney, synovium, and odontoblasts. TRPC5 has been recently confirmed to play a key role in spontaneous, inflammatory mechanical, and cold pain. Although TRPC5 activation is known to be cold sensitive, it is unclear whether this property is intrinsic to the channel protein and whether or to what extent it may be determined by the cellular environment. In this study, we explored the cold sensitivity of human TRPC5 at the single-channel level using transiently transfected HEK293T cells. Upon decreasing the temperature, the channel demonstrated prolonged mean open dwell times and a robust increase in the open probability (P-o), whereas the amplitude of unitary currents decreased similar to 1.5-fold per 10 degrees C of temperature difference. In the absence of any agonists, the temperature dependence of P-o was sigmoidal, with a steep slope within the temperature range of 16 degrees C-11 degrees C, and exhibited saturation below 8-5 degrees C. Thermodynamic analysis revealed significant changes in enthalpy and entropy, suggesting that substantial conformational changes accompany cold-induced gating. The mutant channel T970A, in which the regulation downstream of G-protein coupled receptor signaling was abrogated, exhibited higher basal activity at room temperature and a less steep temperature response profile, with an apparent threshold below 22 degrees C. An even more pronounced decrease in the activation threshold was observed in a mutant that disrupted the electrostatic interaction of TRPC5 with the endoplasmic reticulum calcium sensor stromal interaction molecule 1. Thus, TRPC5 exhibits features of an intrinsically cold-gated channel, its sensitivity to cold tightly depends on the phosphorylation status of the protein and intracellular calcium homeostasis.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

30103 - Neurosciences (including psychophysiology)

Projekt

<a href="/cs/project/GA22-13750S" target="_blank" >GA22-13750S: Signální dráhy ovlivňující funkci lidského TRPC5 receptoru: predikce jejich vztahu k bolesti při revmatoidní artritidě</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2022

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Journal of Cellular Physiology

ISSN

0021-9541

e-ISSN

1097-4652

Svazek periodika

237

Číslo periodika v rámci svazku

9

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

13

Strana od-do

3614-3626

Kód UT WoS článku

000816966300001

EID výsledku v databázi Scopus

2-s2.0-85132880697